title: Dissecting genealogy and cell cycle as sources of cell-to-cell variability in MAPK signaling using high-throughput lineage tracking.
abstract: Cells, even those having identical genotype, exhibit variability in their response to external stimuli. This variability arises from differences in the abundance, localization, and state of cellular components. Such nongenetic differences are likely heritable between successive generations and can also be influenced by processes such as cell cycle, age, or interplay between different pathways. To address the contribution of nongenetic heritability and cell cycle in cell-to-cell variability we developed a high-throughput and fully automated microfluidic platform that allows for concurrent measurement of gene expression, cell-cycle periods, age, and lineage information under a large number of temporally changing medium conditions and using multiple strains. We apply this technology to examine the role of nongenetic inheritance in cell heterogeneity of yeast pheromone signaling. Our data demonstrate that the capacity to respond to pheromone is passed across generations and that the strength of the response correlations between related cells is affected by perturbations in the signaling pathway. We observe that a ste50Δ mutant strain exhibits highly heterogeneous response to pheromone originating from a unique asymmetry between mother and daughter response. On the other hand, fus3Δ cells were found to exhibit an unusually high correlation between mother and daughter cells that arose from a combination of extended cell-cycle periods of fus3Δ mothers, and decreased cell-cycle modulation of the pheromone pathway. Our results contribute to the understanding of the origins of cell heterogeneity and demonstrate the importance of automated platforms that generate single-cell data on several parameters.
authors.affiliation: Centre for High-Throughput Biology and Department of Physics and Astronomy, University of British Columbia, Vancouver, BC, Canada V6T 1Z4.
authors.forename: Marketa
authors.initials: M
authors.lastname: Ricicova
authors.forename: Mani
authors.initials: M
authors.lastname: Hamidi
authors.forename: Adam
authors.initials: A
authors.lastname: Quiring
authors.forename: Antti
authors.initials: A
authors.lastname: Niemistö
authors.forename: Eldon
authors.initials: E
authors.lastname: Emberly
authors.forename: Carl L
authors.initials: CL
authors.lastname: Hansen
mesh_terms: D002453:Cell Cycle; D019070:Cell Lineage; D015870:Gene Expression; D020935:MAP Kinase Signaling System; D044085:Microfluidics; D012441:Saccharomyces cerevisiae / Q000166:cytology* / Q000201:enzymology
keywords: image analysis; microscopy; single cell